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Details of the target gene |
Gene Model of Rodent Parasite |
PBANKA_1432200
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Gene Model P. falciparum ortholog |
PF3D7_1216500
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Gene product | male development gene 1 | protein of early gametocyte 3 |
Gene product: Alternative name | MDV-1; PEG3; MDV-1/PEG3 |
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Details of the genetic modification |
Name of the tag | BirA*::3xMyc |
Details of tagging | C-terminal |
Additional remarks: tagging | |
Commercial source of tag-antibodies | |
Type of plasmid/construct | (Linear) plasmid single cross-over |
PlasmoGEM (Sanger) construct/vector used | No |
Modified PlasmoGEM construct/vector used | No
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Plasmid/construct map |
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Plasmid/construct sequence |
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Restriction sites to linearize plasmid |
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Selectable marker used to select the mutant parasite | hdhfr |
Promoter of the selectable marker | pbdhfr |
Selection (positive) procedure | pyrimethamine |
Selection (negative) procedure | No |
Additional remarks genetic modification | The 3’ end 661 bps of the MDV1/PEG3 (PBANKA_143220) ORF were PCR-amplified as 2 adjacent products with primers g3094 and g3095, and g3096 and g3097, digested with EcoRI, ligated, and re-amplified with primers g3094 and g3096, thus introducing a unique EcoRI-site in the middle of the amplicon. After digestion with SspI and BamHI the PCR product was ligated in frame with BirA*-cmyc into a plasmid containing the human DHFR selection marker. Prior to transfection the plasmid pLIS0297 was linearised with EcoRI. |
Additional remarks selection procedure | |
Primer information: Primers used for amplification of the target sequences
Primer information: Primers used for amplification of the target sequences
Sequence Primer 1 | |
Additional information primer 1 | |
Sequence Primer 2 | |
Additional information primer 2 | |
Sequence Primer 3 | |
Additional information primer 3 | |
Sequence Primer 4 | |
Additional information primer 4 | |
Sequence Primer 5 | |
Additional information primer 5 | |
Sequence Primer 6 | |
Additional information primer 6 | |
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